One-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human enamelysin (MMP-20)

Ting Wang, Takanori Aoki, Kazushi Iwata, Takashi Takata, Takashi Uchida, Vera Knäuper, Elena Llano, Yasunori Okada, John D. Bartlett

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 20 (MMP-20, enamelysin) was established by use of a solid-phase monoclonal antibody and a separate peroxidase-labeled monoclonal antibody. The EIA system was shown to be sensitive and quantitative for the detection of MMP-20. As little as 1.0 ng/ml (50 pg/assay) of MMP-20 protein could be reliably detected. The EIA system was linear over a range of 2.5-160 ng/ml (125-8,000 pg/assay), and the EIA system was versatile in that it was capable of detecting with equal sensitivity proMMP-20, active MMP-20, and MMP-20 with COOH-terminal deletions. The EIA system was validated by the successful detection of MMP-20 in the culture medium of Chinese hamster ovary cells (CHO-K1) that were transfected with an MMP-20 expression vector. No MMP-20 was detected in normal human serum, normal saliva, or in selected tumors. However, when recombinant human MMP-20 was added to human saliva, the EIA system did detect quantifiable amounts of the MMP-20, indicating that the system will work within the framework of complex in biological fluids.

Original languageEnglish
Pages (from-to)530-537
Number of pages8
JournalEuropean Journal of Oral Sciences
Volume108
Issue number6
DOIs
Publication statusPublished - 2000 Dec

Keywords

  • Enamelysin
  • MMP-20
  • Monoclonal antibody
  • Sandwich enzyme immunoassay

ASJC Scopus subject areas

  • Dentistry(all)

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    Wang, T., Aoki, T., Iwata, K., Takata, T., Uchida, T., Knäuper, V., Llano, E., Okada, Y., & Bartlett, J. D. (2000). One-step sandwich enzyme immunoassay using monoclonal antibodies for detection of human enamelysin (MMP-20). European Journal of Oral Sciences, 108(6), 530-537. https://doi.org/10.1034/j.1600-0722.2000.00922.x